EP0915908A1 - Dolastatin pentapeptide derivatives and their use as antitumor agents - Google Patents
Dolastatin pentapeptide derivatives and their use as antitumor agentsInfo
- Publication number
- EP0915908A1 EP0915908A1 EP97934502A EP97934502A EP0915908A1 EP 0915908 A1 EP0915908 A1 EP 0915908A1 EP 97934502 A EP97934502 A EP 97934502A EP 97934502 A EP97934502 A EP 97934502A EP 0915908 A1 EP0915908 A1 EP 0915908A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- nhch
- group
- och
- compound
- methyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 229930188854 dolastatin Natural products 0.000 title description 5
- AMRJKAQTDDKMCE-UHFFFAOYSA-N dolastatin Chemical compound CC(C)C(N(C)C)C(=O)NC(C(C)C)C(=O)N(C)C(C(C)C)C(OC)CC(=O)N1CCCC1C(OC)C(C)C(=O)NC(C=1SC=CN=1)CC1=CC=CC=C1 AMRJKAQTDDKMCE-UHFFFAOYSA-N 0.000 title description 3
- 239000002246 antineoplastic agent Substances 0.000 title description 2
- -1 2-t-butylglycyl Chemical group 0.000 claims abstract description 38
- 150000001875 compounds Chemical class 0.000 claims abstract description 36
- 238000000034 method Methods 0.000 claims abstract description 26
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 20
- 229950003188 isovaleryl diethylamide Drugs 0.000 claims abstract description 12
- 241000124008 Mammalia Species 0.000 claims abstract description 11
- 125000000539 amino acid group Chemical group 0.000 claims abstract description 10
- 125000003277 amino group Chemical group 0.000 claims abstract description 10
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims abstract description 8
- 239000002253 acid Substances 0.000 claims abstract description 6
- 125000000741 isoleucyl group Chemical group [H]N([H])C(C(C([H])([H])[H])C([H])([H])C([H])([H])[H])C(=O)O* 0.000 claims abstract description 6
- 125000001909 leucine group Chemical group [H]N(*)C(C(*)=O)C([H])([H])C(C([H])([H])[H])C([H])([H])[H] 0.000 claims abstract description 4
- 150000003839 salts Chemical class 0.000 claims abstract description 4
- 125000002987 valine group Chemical group [H]N([H])C([H])(C(*)=O)C([H])(C([H])([H])[H])C([H])([H])[H] 0.000 claims abstract 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 9
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 8
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 7
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 6
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 5
- 239000004305 biphenyl Substances 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 208000026310 Breast neoplasm Diseases 0.000 claims description 3
- VUWZPRWSIVNGKG-UHFFFAOYSA-N fluoromethane Chemical compound F[CH2] VUWZPRWSIVNGKG-UHFFFAOYSA-N 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- 208000029742 colonic neoplasm Diseases 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 208000037841 lung tumor Diseases 0.000 claims description 2
- 206010006187 Breast cancer Diseases 0.000 claims 1
- 150000001412 amines Chemical class 0.000 claims 1
- 125000000753 cycloalkyl group Chemical group 0.000 claims 1
- 208000020816 lung neoplasm Diseases 0.000 claims 1
- 125000004573 morpholin-4-yl group Chemical group N1(CCOCC1)* 0.000 claims 1
- 125000001113 thiadiazolyl group Chemical group 0.000 claims 1
- 125000003258 trimethylene group Chemical group [H]C([H])([*:2])C([H])([H])C([H])([H])[*:1] 0.000 claims 1
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 16
- 239000000203 mixture Substances 0.000 abstract description 15
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 12
- 230000000259 anti-tumor effect Effects 0.000 abstract description 5
- 201000011510 cancer Diseases 0.000 abstract description 5
- 230000036210 malignancy Effects 0.000 abstract description 4
- 125000000217 alkyl group Chemical group 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 40
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 33
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Chemical compound CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 30
- 230000015572 biosynthetic process Effects 0.000 description 25
- 238000003786 synthesis reaction Methods 0.000 description 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 21
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 21
- 239000000243 solution Substances 0.000 description 21
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 15
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 13
- 229940093499 ethyl acetate Drugs 0.000 description 13
- 235000019439 ethyl acetate Nutrition 0.000 description 13
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 12
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 12
- 239000007787 solid Substances 0.000 description 12
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 10
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 10
- 239000011541 reaction mixture Substances 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 9
- 238000003756 stirring Methods 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 241000699670 Mus sp. Species 0.000 description 8
- 230000008878 coupling Effects 0.000 description 8
- 238000010168 coupling process Methods 0.000 description 8
- 238000005859 coupling reaction Methods 0.000 description 8
- 229920006395 saturated elastomer Polymers 0.000 description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 7
- 150000001413 amino acids Chemical class 0.000 description 7
- 239000012043 crude product Substances 0.000 description 7
- OFDNQWIFNXBECV-VFSYNPLYSA-N dolastatin 10 Chemical compound CC(C)[C@H](N(C)C)C(=O)N[C@@H](C(C)C)C(=O)N(C)[C@@H]([C@@H](C)CC)[C@H](OC)CC(=O)N1CCC[C@H]1[C@H](OC)[C@@H](C)C(=O)N[C@H](C=1SC=CN=1)CC1=CC=CC=C1 OFDNQWIFNXBECV-VFSYNPLYSA-N 0.000 description 7
- 239000012634 fragment Substances 0.000 description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- OFDNQWIFNXBECV-UHFFFAOYSA-N Dolastatin 10 Natural products CC(C)C(N(C)C)C(=O)NC(C(C)C)C(=O)N(C)C(C(C)CC)C(OC)CC(=O)N1CCCC1C(OC)C(C)C(=O)NC(C=1SC=CN=1)CC1=CC=CC=C1 OFDNQWIFNXBECV-UHFFFAOYSA-N 0.000 description 6
- 108010045524 dolastatin 10 Proteins 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- ZCSHNCUQKCANBX-UHFFFAOYSA-N lithium diisopropylamide Chemical compound [Li+].CC(C)[N-]C(C)C ZCSHNCUQKCANBX-UHFFFAOYSA-N 0.000 description 6
- 238000010647 peptide synthesis reaction Methods 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 238000004992 fast atom bombardment mass spectroscopy Methods 0.000 description 5
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 5
- 235000019341 magnesium sulphate Nutrition 0.000 description 5
- 229910052938 sodium sulfate Inorganic materials 0.000 description 5
- 235000011152 sodium sulphate Nutrition 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 4
- 150000001408 amides Chemical class 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000004440 column chromatography Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- OAYLNYINCPYISS-UHFFFAOYSA-N ethyl acetate;hexane Chemical compound CCCCCC.CCOC(C)=O OAYLNYINCPYISS-UHFFFAOYSA-N 0.000 description 4
- 125000001500 prolyl group Chemical group [H]N1C([H])(C(=O)[*])C([H])([H])C([H])([H])C1([H])[H] 0.000 description 4
- 125000006239 protecting group Chemical group 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 125000002114 valyl group Chemical group 0.000 description 4
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 3
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 3
- 230000005526 G1 to G0 transition Effects 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 239000003054 catalyst Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 239000012230 colorless oil Substances 0.000 description 3
- 238000002425 crystallisation Methods 0.000 description 3
- 230000008025 crystallization Effects 0.000 description 3
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- GNOIPBMMFNIUFM-UHFFFAOYSA-N hexamethylphosphoric triamide Chemical compound CN(C)P(=O)(N(C)C)N(C)C GNOIPBMMFNIUFM-UHFFFAOYSA-N 0.000 description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 3
- YNESATAKKCNGOF-UHFFFAOYSA-N lithium bis(trimethylsilyl)amide Chemical compound [Li+].C[Si](C)(C)[N-][Si](C)(C)C YNESATAKKCNGOF-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000012044 organic layer Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- CHKVPAROMQMJNQ-UHFFFAOYSA-M potassium bisulfate Chemical compound [K+].OS([O-])(=O)=O CHKVPAROMQMJNQ-UHFFFAOYSA-M 0.000 description 3
- 229910000343 potassium bisulfate Inorganic materials 0.000 description 3
- 238000002953 preparative HPLC Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- JVSFQJZRHXAUGT-UHFFFAOYSA-N 2,2-dimethylpropanoyl chloride Chemical compound CC(C)(C)C(Cl)=O JVSFQJZRHXAUGT-UHFFFAOYSA-N 0.000 description 2
- 241000237373 Aplysia sp. Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- 108010016626 Dipeptides Proteins 0.000 description 2
- 241000237378 Dolabella auricularia Species 0.000 description 2
- LQKSHSFQQRCAFW-UHFFFAOYSA-N Dolastatin 15 Natural products COC1=CC(=O)N(C(=O)C(OC(=O)C2N(CCC2)C(=O)C2N(CCC2)C(=O)C(C(C)C)N(C)C(=O)C(NC(=O)C(C(C)C)N(C)C)C(C)C)C(C)C)C1CC1=CC=CC=C1 LQKSHSFQQRCAFW-UHFFFAOYSA-N 0.000 description 2
- 239000007821 HATU Substances 0.000 description 2
- 208000002250 Hematologic Neoplasms Diseases 0.000 description 2
- 208000028018 Lymphocytic leukaemia Diseases 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- APGLTERDKORUHK-LURJTMIESA-N N,N-dimethyl-L-Valine Chemical compound CC(C)[C@H](N(C)C)C(O)=O APGLTERDKORUHK-LURJTMIESA-N 0.000 description 2
- LQKSHSFQQRCAFW-CCVNJFHASA-N [(2s)-1-[(2s)-2-benzyl-3-methoxy-5-oxo-2h-pyrrol-1-yl]-3-methyl-1-oxobutan-2-yl] (2s)-1-[(2s)-1-[(2s)-2-[[(2s)-2-[[(2s)-2-(dimethylamino)-3-methylbutanoyl]amino]-3-methylbutanoyl]-methylamino]-3-methylbutanoyl]pyrrolidine-2-carbonyl]pyrrolidine-2-carboxyl Chemical compound C([C@@H]1N(C(=O)C=C1OC)C(=O)[C@@H](OC(=O)[C@H]1N(CCC1)C(=O)[C@H]1N(CCC1)C(=O)[C@H](C(C)C)N(C)C(=O)[C@@H](NC(=O)[C@H](C(C)C)N(C)C)C(C)C)C(C)C)C1=CC=CC=C1 LQKSHSFQQRCAFW-CCVNJFHASA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000000118 anti-neoplastic effect Effects 0.000 description 2
- 239000008346 aqueous phase Substances 0.000 description 2
- SHJZPYCTHTUGQA-AWEZNQCLSA-N benzyl (2s)-2-(propan-2-ylcarbamoyl)pyrrolidine-1-carboxylate Chemical compound CC(C)NC(=O)[C@@H]1CCCN1C(=O)OCC1=CC=CC=C1 SHJZPYCTHTUGQA-AWEZNQCLSA-N 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 229940126214 compound 3 Drugs 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- VLNZUSMTOFYNPS-UHFFFAOYSA-N diethylphosphorylformonitrile Chemical compound CCP(=O)(CC)C#N VLNZUSMTOFYNPS-UHFFFAOYSA-N 0.000 description 2
- MKRTXPORKIRPDG-UHFFFAOYSA-N diphenylphosphoryl azide Chemical compound C=1C=CC=CC=1P(=O)(N=[N+]=[N-])C1=CC=CC=C1 MKRTXPORKIRPDG-UHFFFAOYSA-N 0.000 description 2
- 108010045552 dolastatin 15 Proteins 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000003966 growth inhibitor Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- 208000003747 lymphoid leukemia Diseases 0.000 description 2
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- UBQKCCHYAOITMY-UHFFFAOYSA-N pyridin-2-ol Chemical compound OC1=CC=CC=N1 UBQKCCHYAOITMY-UHFFFAOYSA-N 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- XACLOOAIJNCRDI-PWSUYJOCSA-N tert-butyl (3r,4s)-3-methoxy-5-methyl-4-(methylamino)hexanoate Chemical compound CN[C@@H](C(C)C)[C@H](OC)CC(=O)OC(C)(C)C XACLOOAIJNCRDI-PWSUYJOCSA-N 0.000 description 2
- RHSLHKPSUHBBTQ-MJGOQNOKSA-N tert-butyl (3r,4s)-3-methoxy-5-methyl-4-[methyl(phenylmethoxycarbonyl)amino]hexanoate Chemical compound CC(C)(C)OC(=O)C[C@@H](OC)[C@H](C(C)C)N(C)C(=O)OCC1=CC=CC=C1 RHSLHKPSUHBBTQ-MJGOQNOKSA-N 0.000 description 2
- FGKFTCDIARXOCS-KRWDZBQOSA-N tert-butyl (4s)-5-methyl-3-oxo-4-(phenylmethoxycarbonylamino)hexanoate Chemical compound CC(C)(C)OC(=O)CC(=O)[C@H](C(C)C)NC(=O)OCC1=CC=CC=C1 FGKFTCDIARXOCS-KRWDZBQOSA-N 0.000 description 2
- 229940086542 triethylamine Drugs 0.000 description 2
- 239000004474 valine Substances 0.000 description 2
- XGDABHXCVGCHBB-QWRGUYRKSA-N (2s)-1-[(2s)-1-[(2-methylpropan-2-yl)oxycarbonyl]pyrrolidine-2-carbonyl]pyrrolidine-2-carboxylic acid Chemical compound CC(C)(C)OC(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 XGDABHXCVGCHBB-QWRGUYRKSA-N 0.000 description 1
- JXGVXCZADZNAMJ-NSHDSACASA-N (2s)-1-phenylmethoxycarbonylpyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1=CC=CC=C1 JXGVXCZADZNAMJ-NSHDSACASA-N 0.000 description 1
- IXSYUULNYWPWNY-ZCFIWIBFSA-N (2s)-2-(dimethylamino)-3,3-dimethylbutanoic acid Chemical compound CN(C)[C@H](C(O)=O)C(C)(C)C IXSYUULNYWPWNY-ZCFIWIBFSA-N 0.000 description 1
- OQJJVXKMPUJFJK-BQBZGAKWSA-N (2s,3s)-2-(dimethylamino)-3-methylpentanoic acid Chemical compound CC[C@H](C)[C@H](N(C)C)C(O)=O OQJJVXKMPUJFJK-BQBZGAKWSA-N 0.000 description 1
- 125000000229 (C1-C4)alkoxy group Chemical group 0.000 description 1
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- RAIPHJJURHTUIC-UHFFFAOYSA-N 1,3-thiazol-2-amine Chemical compound NC1=NC=CS1 RAIPHJJURHTUIC-UHFFFAOYSA-N 0.000 description 1
- FNQIGYRDLYROLW-UHFFFAOYSA-N 1-hydroxy-2h-1,2,3-benzotriazine Chemical compound C1=CC=C2N(O)NN=CC2=C1 FNQIGYRDLYROLW-UHFFFAOYSA-N 0.000 description 1
- BSXPDVKSFWQFRT-UHFFFAOYSA-N 1-hydroxytriazolo[4,5-b]pyridine Chemical compound C1=CC=C2N(O)N=NC2=N1 BSXPDVKSFWQFRT-UHFFFAOYSA-N 0.000 description 1
- MQLACMBJVPINKE-UHFFFAOYSA-N 10-[(3-hydroxy-4-methoxyphenyl)methylidene]anthracen-9-one Chemical compound C1=C(O)C(OC)=CC=C1C=C1C2=CC=CC=C2C(=O)C2=CC=CC=C21 MQLACMBJVPINKE-UHFFFAOYSA-N 0.000 description 1
- NBWRJAOOMGASJP-UHFFFAOYSA-N 2-(3,5-diphenyl-1h-tetrazol-1-ium-2-yl)-4,5-dimethyl-1,3-thiazole;bromide Chemical compound [Br-].S1C(C)=C(C)N=C1N1N(C=2C=CC=CC=2)N=C(C=2C=CC=CC=2)[NH2+]1 NBWRJAOOMGASJP-UHFFFAOYSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- WDYVUKGVKRZQNM-UHFFFAOYSA-N 6-phosphonohexylphosphonic acid Chemical compound OP(O)(=O)CCCCCCP(O)(O)=O WDYVUKGVKRZQNM-UHFFFAOYSA-N 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- 206010066476 Haematological malignancy Diseases 0.000 description 1
- 125000003580 L-valyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(C([H])([H])[H])(C([H])([H])[H])[H] 0.000 description 1
- 241001024304 Mino Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000011717 athymic nude mouse Methods 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- CCAFPWNGIUBUSD-UHFFFAOYSA-N diethyl sulfoxide Chemical compound CCS(=O)CC CCAFPWNGIUBUSD-UHFFFAOYSA-N 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- MHPUGCYGQWGLJL-UHFFFAOYSA-N dimethyl pentanoic acid Natural products CC(C)CCCC(O)=O MHPUGCYGQWGLJL-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 208000023965 endometrium neoplasm Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- JBFYUZGYRGXSFL-UHFFFAOYSA-N imidazolide Chemical compound C1=C[N-]C=N1 JBFYUZGYRGXSFL-UHFFFAOYSA-N 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000005462 in vivo assay Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- OIRDBPQYVWXNSJ-UHFFFAOYSA-N methyl trifluoromethansulfonate Chemical compound COS(=O)(=O)C(F)(F)F OIRDBPQYVWXNSJ-UHFFFAOYSA-N 0.000 description 1
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- XCVNDBIXFPGMIW-UHFFFAOYSA-N n-ethylpropan-1-amine Chemical compound CCCNCC XCVNDBIXFPGMIW-UHFFFAOYSA-N 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 238000011580 nude mouse model Methods 0.000 description 1
- 230000000771 oncological effect Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 description 1
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl chloride Substances ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 208000023958 prostate neoplasm Diseases 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- DCKVNWZUADLDEH-UHFFFAOYSA-N sec-butyl acetate Chemical compound CCC(C)OC(C)=O DCKVNWZUADLDEH-UHFFFAOYSA-N 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 239000007885 tablet disintegrant Substances 0.000 description 1
- ASBBSKISGIQILH-WBVHZDCISA-N tert-butyl (3r,4s)-3-hydroxy-5-methyl-4-(phenylmethoxycarbonylamino)hexanoate Chemical compound CC(C)(C)OC(=O)C[C@@H](O)[C@H](C(C)C)NC(=O)OCC1=CC=CC=C1 ASBBSKISGIQILH-WBVHZDCISA-N 0.000 description 1
- WMOVHXAZOJBABW-UHFFFAOYSA-N tert-butyl acetate Chemical compound CC(=O)OC(C)(C)C WMOVHXAZOJBABW-UHFFFAOYSA-N 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000001544 thienyl group Chemical group 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/02—Linear peptides containing at least one abnormal peptide link
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- Dolastatins 10 and 15 are the most potent cell growth inhibitors. Dolastatin 15, for example, inhibits the growth of the National Cancer Institute's
- P388 lymphocytic leukemia (PS system) cell line a string predictor of efficacy against various types of human malignancies.
- Dolastatin 10 and Dolastatin 15 effectively inhibit tubulin polymerization and growth of four different human lymphoma cell lines (Bai et al., Biochem. Pharmacol. 39: 1941-1949 (1990); 0 Beckwith et al., supra (1993)).
- Dolastatin 10 Pettit et al., J. Am. Chem. Soc. Ill: 5463-5465 (1989); Roux et al . Tetrahedron 50: 5345-5360 (1994); Shiori et al. Tetrahedron 49: 1913-1924 (1993)).
- Synthetic Dolastatin 10, 0 suffers from disadvantages which include poor solubility in aqueous systems and the need for expensive starting materials for its synthesis.
- the present invention provides peptides which have antitumor or anti-neoplastic activity. These compounds are of Formula I,
- A is an amino acid residue of the formula (CH 3 ) 2 N-CHX-CO, wherein X is a normal or branched C 3 . 4 -alkyl group.
- B is an amino acid residue selected form the group consisting of valyl, isoleucyl, leucyl, and 2- -butylglycyl .
- D is a normal or branched C 3 -C 4 -alkyl group.
- K is a t-butoxy group or a substituted amino group.
- compositions comprising a compound of Formula I and a pharmaceutically acceptable carrier.
- a additional embodiment of the present invention is a method for treating a malignancy in a mammal, such as a human, comprising administering to the mammal an effective amount of a compound or compounds of Formula I in a pharmaceutically acceptable composition.
- the present invention relates to peptides having antitumor or antineoplastic activity. It also includes pharmaceutical compositions comprising these compounds and methods for treating cancer in a mammal, including a human, by administration of these compositions to the mammal.
- Dolastatin 10 results in novel compounds with a surprisingly improved therapeutic potential for the treatment of neoplastic diseases, as compared to Dolastatin 10. Furthermore, the compound of the present invention can be conveniently synthesized, as described below in detail .
- Compounds of the present invention include antitumor peptides of Formula I:
- A is an amino acid residue of the formula (CH 3 ) 2 N-CHX-C0, wherein X is a normal or branched C . 4 -alkyl group;
- B is an amino acid residue selected from the group consisting of valyl, isoleucyl, leucyl, and 2 - t-butylglycyl ;
- D is a normal or branched C 3-4 -alkyl group; and
- K is a t-butoxy group or a substituted amino group.
- Suitable amino groups include: -N(C ⁇ - 3 -alkyl)C ⁇ - 3 -alkyl, normal or branched -NH-Ci- ⁇ -alkyl, -NH-C(CH 3 ) 2 CN, -NH-C(CH 3 ) 2 CCH, -NH-C (CH 3 ) 2 CH 2 CH 2 0H, -NH-C(CH 3 ) 2 CH 2 OH, -NH-C 3 .
- Preferred compounds of the present invention are of Formula I in which:
- A is an amino acid residue of the formula (CH 3 ) 2N-CHX-CO, wherein X is an isopropyl, t-butyl or sec-butyl group;
- B is an amino acid residue selected from the group consisting of valyl, isoleucyl and 2- t-butylglycyl;
- D is an isopropyl, t-butyl or sec-butyl group
- K is a t-butoxy group or a substituted amino group selected from among the following: -NHCH 3 , -NHCH 2 CH 3 , -NH(CH 2 ) 2 CH 3 , -NH(CH 2 ) 3 CH 3 ,
- -NH-adamantyl (2) -NH-adamantyl (1) , -NH-CH 2 -naphthyl, -NH-benz- hydryl, -NH-biphenyl, -NH-pyridyl, -NH-CH 2 -pyridyl, -NH-CH 2 -CH 2 - pyridyl, -NH-benzothiazolyl, -NH-benzoisothiazolyl, -NH-benzo- pyrazolyl, -NH-benzoxazolyl, -NH-fluorenyl, -NH-pyrimidyl, -NH-CH 2 - (4-methyl) thiazolyl (2) , -NH-CH 2 -furanyl (2) ,
- the compounds of the present invention can be prepared by known methods of peptide synthesis.
- the peptides can be assembled sequentially from individual amino acids or by linking suitable small peptide fragments.
- sequential assembly the peptide chain is extended stepwise, starting at the C- terminus, by one amino acid per step.
- fragment coupling fragments of different lengths can be linked together, and the fragments in turn can be obtained by sequential assembly from amino acids or by fragment coupling of still shorter peptides.
- Preferred methods include the azide method, the syrnrnetric and mixed anhydride method, the use of in situ generated or preformed active esters, the use of urethane protected N-carboxy anhydrides of amino acids and the formation of the amide linkage using coupling reagents, such as carboxylic acid activators, especially dicyclohexylcarbodiimide (DCC) , diisopropylcarbodii ide (DIC) , l-ethoxycarbonyl-2-ethoxy-l,2-dihydroquinoline (EEDQ) , l-ethyl-3- (3 -dimethylaminopropyDcarbodiimide hydrochloride (EDCI) , n-propane-phosphonic anhydride (PPA) , N,N- bis (2-oxo-oxazolidi- nyl) imido-phosphoryl chloride (B0P-C1) , bromo- tris-pyrrolidin
- the coupling reagents can be employed alone or in combination with additives such as N,N-dimethyl-4 -amino- pyridine (DMAP) , N-hydroxybenzotriazole (HOBt) , N-hydroxybenzo- triazine (HOOBt), N-hydroxyazabenzotriazole (HOAt) , N-hydroxy- succinimide (HOSu) or 2-hydroxypyridine.
- DMAP N,N-dimethyl-4 -amino- pyridine
- HOBt N-hydroxybenzotriazole
- HOOBt N-hydroxybenzo- triazine
- HOAt N-hydroxyazabenzotriazole
- HSu N-hydroxy- succinimide
- 2-hydroxypyridine 2-hydroxypyridine
- protecting groups are generally not necessary in enzymatic peptide synthesis, reversible protection of reactive groups not involved in formation of the amide linkage is necessary for both reactants in chemical synthesis.
- Three conventional protective group techniques are preferred for chemical peptide synthesis: the benzyloxycarbonyl (Z) , the t-butoxy- carbonyl (Boc) and the 9-fluorenylmethoxycarbonyl (Fmoc) techniques. Identified in each case is the protective group on the ⁇ -amino group of the chain-extending unit.
- a detailed review of amino-acid protective groups is given by M ⁇ ller, Methoden der organischen Chemie Vol.
- Solvents suitable for peptide synthesis include any solvent which is inert under the reaction conditions, especially water, N,N-dimethylformamide (DMF) , dimethyl sulfoxide (DMSO) , aceto- nitrile, dichloromethane (DCM) , 1,4-dioxane, tetrahydrofuran (THF) , N-methyl-2-pyrrolidone (NMP) , ethyl acetate and mixtures of these solvents.
- DMF N,N-dimethylformamide
- DMSO dimethyl sulfoxide
- DCM dichloromethane
- THF tetrahydrofuran
- NMP N-methyl-2-pyrrolidone
- NCAs BOC-protected amino acid N-carboxy anhydrides
- Z-protected NCAs the use of pivaloyl chloride or HATU as the condensing agent is most advantageous for this type of coupling.
- Peptides which are dialkylated at the amino terminus can be prepared using the appropriate N,N-dialkylamino acid as a building block or by hydrogenating N-unsubstituted peptides in solution in the presence of an appropriate aldehyde or ketone and a catalyst such as palladium on charcoal.
- the various non-naturally occurring amino acids disclosed herein can be obtained from commercial sources or synthesized from commercially available materials using methods known in the art.
- the moiety -NR 3 -CHD-CH/OCG 3 ) CH 2 CO- can be prepared according to published procedures (Shiori et al . in Peptide Chemistry, Yanaihara, ed. (1989); Pettit et al . , J. Am. Chem. Soc. Ill: 5463 (1989); Shiori et al . , Tet. Lett. 931-934 (1991); Koga et al., Tet. Lett. 2395-2398 (1991)).
- the present invention comprises a method for partially or totally inhibiting formation of, or otherwise treating (e.g., reversing or inhibiting the further development of) solid tumors (e.g., tumors of the lung, breast, colon, prostate, bladder, rectum, or endometrial tumors) or hemato- logical malignancies (e.g., leukemias, lymphomas) in a mammal, for example, a human, by administering to the mammal a thera- Chamberically effective amount of a compound or a combination of compounds of Formula I.
- solid tumors e.g., tumors of the lung, breast, colon, prostate, bladder, rectum, or endometrial tumors
- hemato- logical malignancies e.g., leukemias, lymphomas
- the compoun (s) of Formula I can be administered alone or in conjunction with other drugs, such as other anti -cancer drugs or in a pharmaceutical composition further comprising an acceptable carrier or diluent, and, optionally, other drugs.
- Administration can be by any means which are appropriate for pharmaceutical, preferably oncological, agents, including oral and parenteral means such as subcuta- neously, intravenously, intramuscularly and intraperitoneally, nasally or rectally.
- the dosage to be administered to the mammal, such as a human is a therapeutically effective amount of a compound described herein.
- the therapeutically effective amount can be administered in a single dose or multiple doses in a given period of time (e.g., a single daily dose or two or more doses a day) .
- "therapeutically effective amount” is an amount sufficient to inhibit (partially or totally) formation of a tumor or a hematological malignancy or to reverse development of a solid tumor or other malignancy or prevent or reduce its further progression.
- the dosage is determined empirically, using known methods, and will depend upon factors such as the biological activity of the particular compound employed; the means of the recipient; the nature and extent of the symptoms; the frequency of treatment; the administration of other therapies; and the effect desired.
- a typical daily dose will be from about 0.5 to about 50 milligrams per kilogram of body weight by oral administration and from about 0.05 to about 20 milligrams per kilogram of body weight by parenteral administration.
- the compounds of the present invention can be administered in conventional solid or liquid pharmaceutical administration forms, for example, uncoated or (film-) coated tablets, capsules, powders, granules, suppositories or solutions. These are produced in a conventional manner.
- the active substances can, for this purpose, be processes with conventional pharmaceutical aids such as tablet binders fillers, preservatives, tablet disintegrants, flow regulators, plasticizers, wetting agents, dispersants, emulsifiers, solvents, sustained release compositions, anti- oxidants and/or propellant gases (cf. H. Sucker et al . : Pharma- zeutician Technologie, Thieme-Verlag, Stuttgart, 1978).
- the administration forms obtained in this way typically contain from about 1 to about 90% by weight of the active substance.
- DCM dichloromethane
- DMSO di ethylsulfoxide
- Bu butyl
- Et ethyl
- Me methyl
- Bzl benzyl
- LDA lithium diisopropylamide
- LHMDS lithium hexamethyldisilazide
- HMPA hexamethylphosphoric triamide
- the purity of the resulting products was determined by analytical HPLC (stationary phase: 100 2.1 mm VYDAC C-18, 10 300 Angstrom; mobile phase: acetonitrile-water gradient, buffered with 0.1% TFA, 40%C) . Characterization was by mass spectroscopy (ESI or FAC-MS) .
- reaction mixture was diluted with saturated aqueous NaHC0 3 (20 ml) was washed with toluene/ethyl - acetate 2:1 (3x) .
- the combined organic layers were extracted with 2 N NCI (3 x 10 ml).
- the aqueous phase was then neutralized with NaHC0 3 an extracted with toluene/ethyl acetate (2:1) (3 x 20 ml).
- the organic layer was dried over magnesium sulfate and concentrated in vacuo.
- Boc-prolyl-proline (1 g, 3.2 mmol) and 2-aminothiazol (0.357 g, 3.2 mmol) were dissolved in 25 ml DMF and cooled to 0°C.
- Triethyl- amine (1.1 ml, 7.6 mmol) was added, followed by DEPC (0.613 ml, 3.7 mmol) .
- the reaction mixture was diluted with ethyl acetate/toluene (2:1), washed with 1 M aqueous potassium hydrogen sulfate, water, saturated aqueous NaHC0 3 and aqueous NaCl, dried over sodium sulfate and concentrated in vacuo.
- the crude product was purified via preparative HPLC to yield 161 mg of the desired product as a white solid.
- N-Benzyloxycarbonyl-prolyl-isopropylamide (28.48 g, 98.2 mmol) was dissolved in 75 ml methanol . This solution was added to a slurry of 3 spatulafuls of 10% Pd/C in 50 ml methanol under a stream of nitrogen. The reaction mixture was hydrogenated for 4 h, filtered over CELITE, washed with methanol, and the solvent evaporated under reduced pressure to give 16 g of a yellow crystalline solid. This material was dissolved in dichloromethane and washed with 5% citric acid (2x) .
- Cytotoxicity was measured using the microculture tetrazolium assay (MTT) , a standard methodology for adherent cell lines. Details of this assay have been published (Alley, et al.. Cancer Research 48: 589-601 (1988)). Exponentially growing cultures of HT-29 colon carcinoma cells were used to make microtiter plate cultures. Cells were seeded at 5000-20,000 cells per well in 96-well plates (in 150 ⁇ l of media), and grown overnight at 37°C. Test compounds were added in 10-fold dilutions varying from 10 " to 10- ⁇ ° M. Cells were then incubated for 48 hours. To determine the number of viable cells in each well, the MTT dye was added
- the concentration of test compound which gives a T/C of 50% growth inhibition was designated as the IC 50 value.
- Compounds of this invention can be tested in a pre- clinical assay for in vivo activity which is indicative of clinical utility.
- Such assays are typically conducted with nude mice into which tumor tissue, preferably of human origin, had been transplanted (xenografted) , as is well known in this field.
- Test compounds are evaluated for anti -tumor efficacy following administration to the xenograft-bearing mice.
- human breast tumors which have been growth in athymic nude mice are transplanted into new recipient mice, using tumor fragments of about 50 mg in size.
- the day of transplantation is designated as day 0.
- the mice are divided into four groups of 5-10 mice each.
- An untreated group serves as the control.
- Doses can be, for example, administered on a schedule such as on days 5, 7, 9, 12, 14, 16, 19, 21 and 23 post -implantation.
- Tumor diameters and body weights are measured twice weekly. Tumor volumes are calculated using the diameters measured with Vernier calipers, and the formula
- Mean tumor volumes are calculated for each treatment group, and T/C values determines for each group relative to the untreated control tumors.
- the P388 murine lymphocytic leukemia is harvested from donor mice by peritoneal lavage at day 7 post- transplant and administered to test mice. Treatment of test mice begins 3 days post-transplant and the drugs are administered for five consecutive days. The survival time for untreated mice is typically between 11 and 13 days. The data are expressed as mean survival time (MST) and tretade/control %. According to National Cancer Institute guidelines, a T/C in the range of 128-190 % indicates a drug with moderate to good activity.
- MST mean survival time
- tretade/control % According to National Cancer Institute guidelines, a T/C in the range of 128-190 % indicates a drug with moderate to good activity.
Abstract
The present invention provides anti-tumor peptides of the Formula (I), A-B-N(CH3)-CHD-CH(OCH3)-CH2CO-Pro-Pro-K, and the acid salts thereof. A is an amino acid residue of the formula (CH3)2N-CHX-CO, wherein X is a normal or branched alkyl group. B is an amino acid residue selected from the group consisting of valyl, isoleucyl, leucyl, and 2-t-butylglycyl. D is a normal or branched C3-C4-alkyl group. K is a t-butoxy group or a substituted amino group. An additional embodiment of the present invention is a method for treating a malignancy in a mammal, such as a human, comprising administering to the mammal an effective amount of a compound or compounds of Formula (I) in a pharmaceutically acceptable composition.
Description
DOLASTATIN PENTAPEPTTDE DERIVATIVES AND THEIR USE AS APΠTTUMOR AGENTS
Background of the Invention 5
A series of short peptides with significant activity as cell growth inhibitors have been isolated from the Indian Ocean sea hare Dolabella auricularia (Pettit et al . , J. Am. Chem. Soc. 109: 6883-6885 (1987); Beckwith et al., J. Natl . Cancer Inst. 85,
10 483-88 (1993); United States Patent No. 4,816,444; European Patent Application Publication No. 398558) . These peptides are referred to as Dolastatins 1-15. Of these, Dolastatins 10 and 15 are the most potent cell growth inhibitors. Dolastatin 15, for example, inhibits the growth of the National Cancer Institute's
15 P388 lymphocytic leukemia (PS system) cell line, a string predictor of efficacy against various types of human malignancies. Dolastatin 10 and Dolastatin 15 effectively inhibit tubulin polymerization and growth of four different human lymphoma cell lines (Bai et al., Biochem. Pharmacol. 39: 1941-1949 (1990); 0 Beckwith et al., supra (1993)).
The minute amounts of the Dolastatin peptides present in Dolabella auricularia (about 1 mg each per 100 kg sea hare) and the consequent difficulties in purifying amounts sufficient 5 for evaluation and use, have motivated efforts toward the synthesis of the more promising of these compounds, including Dolastatin 10 (Pettit et al., J. Am. Chem. Soc. Ill: 5463-5465 (1989); Roux et al . Tetrahedron 50: 5345-5360 (1994); Shiori et al. Tetrahedron 49: 1913-1924 (1993)). Synthetic Dolastatin 10, 0 however, suffers from disadvantages which include poor solubility in aqueous systems and the need for expensive starting materials for its synthesis. These disadvantages, in turn, have led to the synthesis and evaluation of structurally modified Dolastatin 10 derivatives (European Patent Application, Publication No. 5 WO 93/03054; Japanese Patent Application No. 06234790; U.S. Patent Application Serial No. 08/178,529).
A need persists for synthetic compounds with the biological activity of Dolastatin 10 which have useful aqueous solubility 0 and can be produced efficiently and economically.
5
Surtimary of the Invention
The present invention provides peptides which have antitumor or anti-neoplastic activity. These compounds are of Formula I,
A-B-N(CH3) -CHD-CH(OCH3) -CHCO-Pro-Pro-K (I) ,
and include the acid salts thereof . In Formula I
A is an amino acid residue of the formula (CH3) 2N-CHX-CO, wherein X is a normal or branched C3.4 -alkyl group.
B is an amino acid residue selected form the group consisting of valyl, isoleucyl, leucyl, and 2- -butylglycyl .
D is a normal or branched C3-C4-alkyl group.
K is a t-butoxy group or a substituted amino group.
Another aspect of the present invention includes pharmaceutical compositions comprising a compound of Formula I and a pharmaceutically acceptable carrier.
A additional embodiment of the present invention is a method for treating a malignancy in a mammal, such as a human, comprising administering to the mammal an effective amount of a compound or compounds of Formula I in a pharmaceutically acceptable composition.
Detailed Description of the Invention
The present invention relates to peptides having antitumor or antineoplastic activity. It also includes pharmaceutical compositions comprising these compounds and methods for treating cancer in a mammal, including a human, by administration of these compositions to the mammal.
Applicants have discovered that structural modification of Dolastatin 10 results in novel compounds with a surprisingly improved therapeutic potential for the treatment of neoplastic diseases, as compared to Dolastatin 10. Furthermore, the compound of the present invention can be conveniently synthesized, as described below in detail .
Compounds of the present invention include antitumor peptides of Formula I:
A-B-N(CH3) -CHD-CH(OCH3) -CH2CO- Pro- ro- K (I) ,
wherein
A is an amino acid residue of the formula (CH3) 2N-CHX-C0, wherein X is a normal or branched C .4 -alkyl group;
B is an amino acid residue selected from the group consisting of valyl, isoleucyl, leucyl, and 2 - t-butylglycyl ; D is a normal or branched C3-4 -alkyl group; and K is a t-butoxy group or a substituted amino group. Examples of suitable amino groups include: -N(Cι-3-alkyl)Cι-3-alkyl, normal or branched -NH-Ci-β-alkyl, -NH-C(CH3)2CN, -NH-C(CH3)2CCH, -NH-C (CH3) 2CH2CH20H, -NH-C(CH3)2CH2OH, -NH-C3.8-cycloalkyl, -NH- [3, 3, 0] -bicyclooctyl, -NHCH(CH3)CH(0H)C6H5, -H-quinolyl, -NH-pyrazyl, -NH-CH2-benz- imidazolyl, -NH-ada antyl, -NH-CH2-adamantyl, -NH-CH (CH3) -phenyl, -NH-C(CH3)2-phenyl, -N{Cι-4-alkoxy) -Ci-4 -alkyl, -N(Cι- -alkoxy) - CH2-phenyl, -N (C1-4-alkoxy) phenyl, -N (CH3)0-phenyl, -NH- (CH2)V- phenyl (v = 0, 1, 2, or 3), -NH- (CH2)m-naphthyl (m = 0 or 1),
-NH- (CH2)w-t>enzhydryl (w = 0, 1, or 2), -NH-biphenyl, -NH-pyridyl, -NH-CH2-pyridyl, -NH-CH2-CH2-pyridyl, -NH-benzothiazolyl, -NH-benzoisothiazolyl, -NH-benzopyrazolyl, -NH-benzoxazolyl, -NH- (CH2)m-fluorenyl (m = 0 or 1) , -NH-pyri idyl, -NH- (CH2)m- indanyl (m = 0 or 1) , -NH- (CH2CH20)y-CH2CH3 (y = 0, 1, 2, 3, 4, or 5), -NH- (CH2CH20)y-CH3 (y = 0, 1, 2, 3, 4, or 5), -NH-Nri-C6H5, -NH-N(CH3)C6H5, -NH-NH-CH2-C6H5, and -NH-N(CH3) CH2-C6H5. K can also be selected from among the following:
^ f~Λ
■ N BJ — N 0 — N SJ •N — N
\_y ^ 0
ci Br
COOCH3
-
COOCH3 C00CH3
■
H3C CH3 H3C CH3
H3
Preferred compounds of the present invention are of Formula I in which:
A is an amino acid residue of the formula (CH3) 2N-CHX-CO, wherein X is an isopropyl, t-butyl or sec-butyl group;
B is an amino acid residue selected from the group consisting of valyl, isoleucyl and 2- t-butylglycyl;
D is an isopropyl, t-butyl or sec-butyl group;
K is a t-butoxy group or a substituted amino group selected from among the following: -NHCH3, -NHCH2CH3, -NH(CH2)2CH3, -NH(CH2) 3CH3,
-NH(CH2)4CH3, -NH(CH2)5CH3, -NHCH(CH3)2, -NHCH (CH2CH3) 2 ,
-NHCH(CH2CH2CH3)2, -NHC(CH3)3, -NHCH [CH(CH3) ] 7,
-NHCH(CH2CH3)CH2CH2CH3, -NHCH (CH3) CH2CH3, -NHCH2CH2F,
-NHC(CH3)2CH2CH3 -NHCH(CH3) CH (CH3) 2, -NHCH(CH3) C (CH3) 3. -NHCH(CH3)CH2CH2CH3 -NHCH2CH (CH3) 2, -NHCH2C (CH3) 3, -NH-cyclopropyl ,
-NH-cyclobutyl, -NH-cyclopentyl, -NH-cyclohexyl, -NH-cycloheptyl,
-N(CH3)OCH3, -N(CH3)2, -N (CH3) OCH2CH3, -N (CH3)0CH2CH2CH3,
-N(CH3)OCH(CH3)2, -N(CH2CH3)OCH3, -N (CH2CH3) OCH2CH3 ,
-N(CH3)OCH2C6H5, -N(OCH3)CH2-C6H5, -N(CH3) OC6H5, -NH-CH2-C6H5,
-NH(CH2)2C6H5, -NH(CH2)3C6H5, -NHCH (CH3) CH (OH) C6H5, -NH-CH2 -cyclo- hexyl, -NH-indanyl- (1) , -NH-CH2CF3, -NHCH (CH2F) 2, -NHC (CH3) 2CH2OH, -NH(CH2CH20)2CH2CH3, -NHC (CH3) 2CN, -NH-quinolyl, -NH-pyrazyl,
-NH-adamantyl (2) , -NH-adamantyl (1) , -NH-CH2-naphthyl, -NH-benz- hydryl, -NH-biphenyl, -NH-pyridyl, -NH-CH2 -pyridyl, -NH-CH2-CH2- pyridyl, -NH-benzothiazolyl, -NH-benzoisothiazolyl, -NH-benzo- pyrazolyl, -NH-benzoxazolyl, -NH-fluorenyl, -NH-pyrimidyl, -NH-CH2- (4-methyl) thiazolyl (2) , -NH-CH2-furanyl (2) ,
-NH-CH2-thienyl (2) , -NH-CH2- (5-methyl) thienyl (2) , -NH- hiazolyl (2), -NH-isoxazolyl(3) , -NH- (3 -methyl) isoxazolyl (5) , -NH- ( -methyl) isothiazolyl (5) , -NH- (5- trifluoromethyl) hia- diazolyl(2), -NH- (5-cyclopropyl) thiadiazolyl (2) , -NH-(4,5-di- methyl) hiazolyl (2) , -NH- (5-methyl) hiadiazolyl (2) , or K is selected from among the heterocyclic amino groups shown below.
-N , -N O , _N *f
^-J v_y ^o Synthetic Methods
The compounds of the present invention can be prepared by known methods of peptide synthesis. Thus, the peptides can be assembled sequentially from individual amino acids or by linking suitable small peptide fragments. In sequential assembly, the peptide chain is extended stepwise, starting at the C- terminus, by one amino acid per step. In fragment coupling, fragments of different lengths can be linked together, and the fragments in turn can be obtained by sequential assembly from amino acids or by fragment coupling of still shorter peptides.
In both sequential assembly and fragment coupling, it is necessary to link the units by forming an amide linkage, which can be accomplished via a variety of enzymatic and chemical methods. Chemical methods for forming the amide linkage are described in detail in standard references on peptide chemistry, including Mύller, Methoden der organischen Chemie Vol. XV/2, 1-364, Thieme Verlag, Stuttgart, (1974); Stewart and Young, Solid Phase Peptide Synthesis, 31-34 and 71-82, Pierce Chemical Company, Rockford, IL (1984); Bodanszky et al . , Peptide Synthesis, 85-128, John Wiley & Sons, New York, (1976) . Preferred methods include the azide method, the syrnrnetric and mixed
anhydride method, the use of in situ generated or preformed active esters, the use of urethane protected N-carboxy anhydrides of amino acids and the formation of the amide linkage using coupling reagents, such as carboxylic acid activators, especially dicyclohexylcarbodiimide (DCC) , diisopropylcarbodii ide (DIC) , l-ethoxycarbonyl-2-ethoxy-l,2-dihydroquinoline (EEDQ) , l-ethyl-3- (3 -dimethylaminopropyDcarbodiimide hydrochloride (EDCI) , n-propane-phosphonic anhydride (PPA) , N,N- bis (2-oxo-oxazolidi- nyl) imido-phosphoryl chloride (B0P-C1) , bromo- tris-pyrrolidino- phosphonium hexafluorophosphate (PyBrop) , diphenylphosphoryl azide (DPPA) , Castro's reagent (BOP, PyBop) , O-benzotriazolyl- N,N,B', N' -tetramethyluronium salts (HBTU) , O-azabenzotriazolyl- N,N,N' ,N' - tetramethyluronium salts (HATU) , diethylphosphoryl cyanide (DEOCN) , 2, 5-diphenyl-2, 3-dihydro-3-oxo-4-hydroxythio- phene dioxide (Steglich's reagent; GOTDO) , and 1, 1 ' -carbonyl- d imidazole (CDI) . The coupling reagents can be employed alone or in combination with additives such as N,N-dimethyl-4 -amino- pyridine (DMAP) , N-hydroxybenzotriazole (HOBt) , N-hydroxybenzo- triazine (HOOBt), N-hydroxyazabenzotriazole (HOAt) , N-hydroxy- succinimide (HOSu) or 2-hydroxypyridine.
Although the use of protecting groups is generally not necessary in enzymatic peptide synthesis, reversible protection of reactive groups not involved in formation of the amide linkage is necessary for both reactants in chemical synthesis. Three conventional protective group techniques are preferred for chemical peptide synthesis: the benzyloxycarbonyl (Z) , the t-butoxy- carbonyl (Boc) and the 9-fluorenylmethoxycarbonyl (Fmoc) techniques. Identified in each case is the protective group on the α-amino group of the chain-extending unit. A detailed review of amino-acid protective groups is given by Mϋller, Methoden der organischen Chemie Vol. XV/1, pp 20-906, Thieme Verlag, Stuttgart (1974) . The units employed for assembling the peptide chain can be reacted in solution, in suspension or by a method similar to that described by Merrifield, J. Amer. Chem. Soc. 85, 2149 (1963) . Particularly preferred methods are those in which peptides are assembled sequentially or by fragment coupling using the Z, Boc or Fmoc protective group technique.
Solvents suitable for peptide synthesis include any solvent which is inert under the reaction conditions, especially water, N,N-dimethylformamide (DMF) , dimethyl sulfoxide (DMSO) , aceto- nitrile, dichloromethane (DCM) , 1,4-dioxane, tetrahydrofuran (THF) , N-methyl-2-pyrrolidone (NMP) , ethyl acetate and mixtures of these solvents.
For coupling of the amino acid following the N-methylated γ- mino acid derivative, the use of either BOC-protected amino acid N-carboxy anhydrides (NCAs) , Z-protected NCAs or the use of pivaloyl chloride or HATU as the condensing agent is most advantageous for this type of coupling.
Peptides which are dialkylated at the amino terminus can be prepared using the appropriate N,N-dialkylamino acid as a building block or by hydrogenating N-unsubstituted peptides in solution in the presence of an appropriate aldehyde or ketone and a catalyst such as palladium on charcoal.
The various non-naturally occurring amino acids disclosed herein can be obtained from commercial sources or synthesized from commercially available materials using methods known in the art. For example, the moiety -NR3-CHD-CH/OCG3) CH2CO- can be prepared according to published procedures (Shiori et al . in Peptide Chemistry, Yanaihara, ed. (1989); Pettit et al . , J. Am. Chem. Soc. Ill: 5463 (1989); Shiori et al . , Tet. Lett. 931-934 (1991); Koga et al., Tet. Lett. 2395-2398 (1991)).
Methods of Use of the Claimed Compounds
In another embodiment, the present invention comprises a method for partially or totally inhibiting formation of, or otherwise treating (e.g., reversing or inhibiting the further development of) solid tumors (e.g., tumors of the lung, breast, colon, prostate, bladder, rectum, or endometrial tumors) or hemato- logical malignancies (e.g., leukemias, lymphomas) in a mammal, for example, a human, by administering to the mammal a thera- peutically effective amount of a compound or a combination of compounds of Formula I. The compoun (s) of Formula I can be administered alone or in conjunction with other drugs, such as other anti -cancer drugs or in a pharmaceutical composition further comprising an acceptable carrier or diluent, and, optionally, other drugs. Administration can be by any means which are appropriate for pharmaceutical, preferably oncological, agents, including oral and parenteral means such as subcuta- neously, intravenously, intramuscularly and intraperitoneally, nasally or rectally.
The dosage to be administered to the mammal, such as a human, is a therapeutically effective amount of a compound described herein. The therapeutically effective amount can be administered in a single dose or multiple doses in a given period of time (e.g., a single daily dose or two or more doses a day) . As used herein, "therapeutically effective amount" is an amount
sufficient to inhibit (partially or totally) formation of a tumor or a hematological malignancy or to reverse development of a solid tumor or other malignancy or prevent or reduce its further progression. For a particular condition or method of treatment, the dosage is determined empirically, using known methods, and will depend upon factors such as the biological activity of the particular compound employed; the means of the recipient; the nature and extent of the symptoms; the frequency of treatment; the administration of other therapies; and the effect desired. A typical daily dose will be from about 0.5 to about 50 milligrams per kilogram of body weight by oral administration and from about 0.05 to about 20 milligrams per kilogram of body weight by parenteral administration.
The compounds of the present invention can be administered in conventional solid or liquid pharmaceutical administration forms, for example, uncoated or (film-) coated tablets, capsules, powders, granules, suppositories or solutions. These are produced in a conventional manner. The active substances can, for this purpose, be processes with conventional pharmaceutical aids such as tablet binders fillers, preservatives, tablet disintegrants, flow regulators, plasticizers, wetting agents, dispersants, emulsifiers, solvents, sustained release compositions, anti- oxidants and/or propellant gases (cf. H. Sucker et al . : Pharma- zeutische Technologie, Thieme-Verlag, Stuttgart, 1978). The administration forms obtained in this way typically contain from about 1 to about 90% by weight of the active substance.
The following examples are intended to illustrate the invention but are not to be considered limitations of the invention.
Examples
The naturally-occurring amino acids are abbreviated in the examples using the known three- letter code. Other abbreviations employed are: TFA = trifluoroacetic acid; Ac = acetic acid;
DCM = dichloromethane; DMSO = di ethylsulfoxide; Bu = butyl; Et = ethyl; Me = methyl; Bzl = benzyl; LDA = lithium diisopropylamide;
LHMDS = lithium hexamethyldisilazide; HMPA = hexamethylphosphoric triamide.
General Materials and Methods
The compounds of the present invention are synthesized by classical solution synthesis using standard Z- and Boc- ethodology as discussed above.
Purification was carried out by crystallization from the appropriate solvents or mixtures thereof, by medium pressure chroma ography (stationary phase: HD-SIL C-8, 20-45 micron, 100 Angstrom; mobile phase: gradient with A = 0.1% TFA/water, B = 5 0.1% TFA/MeOH), or by preparative HPLC (stationary phase: Waters Delta Pak C-18, 15 micron, 100 Angstrom; mobile phase: gradient with A = 0.1% TFA/water, B = 0.1% TFA(MeOH or 0.1% TFA/Aceto- nitril) . The purity of the resulting products was determined by analytical HPLC (stationary phase: 100 2.1 mm VYDAC C-18, 10 300 Angstrom; mobile phase: acetonitrile-water gradient, buffered with 0.1% TFA, 40%C) . Characterization was by mass spectroscopy (ESI or FAC-MS) .
Example 1 Synthesis Of (3R, 4S) -4- [N- (N,N-dimethyl-L-valyl-L- 15 valyl) -N-methylamino) -3 -methoxy-5-methyl-hexanoyl- prolyl-prolyl-thiazolyl (2) -amide (compound 1)
Synthesis of t-Butyl- (4S) -4- (N-benzyloxycarbonylamino) -5-methyl - 3 -oxohexanoate 0
To an ice-cooled solution of Z-Valine (5 g, 19.9 mmol) in 60 ml tetrahydrofuran was added N,N' -carbonyldiimidazole (3.55 g, 22.3 mmol) in one portion and the resulting mixture stirred for 3 h. t-Butyl acetate (13.5 ml, 100 mmol) was added dropwise to 5 a solution of LDA (90 mmol in tetrahydrofuran (270 ml) at -78°C. After 30 min, the imidazolide solution was added dropwise via double-ended needle to the enolate. The resulting mixture was stirred for 2 h until the temperature rose to -15°C. The reaction was quenched with ether (3 x 200 ml) . The combined organic 0 extracts were washed with 2 N aqueous HC1 (50 ml) , saturated aqueous NaHC03 (2 x 50 ml) , dried over magnesium sulfate and evaporated to dryness in vacuo. The residue was purified by column chromatography on silica gel using ethyl acetate-hexane (1:4) as an eluent to give the product as a colorless oil 5 (6.88 g) .
MS: calc. onoisotopic mass 349.19; found ESI-: 348.1
Synthesis of t -butyl- (3R,4S) -4- (N-benzyloxycarbonylamino) -3- hydroxy- 5 -methy1 -hexanoate 0
To a solution of t-butyl- (4S) -4- (N-benzyloxycarbonylamino) - 5-methyl-3-oxohexanoate (6.0 g, 17.1 mmol) in 70 ml ethanol at 0°C was added potassium borohydride (3.23 g, 58.8 mmol). After stirring for 4 h at 0°C and 12 h at room temperature, the reaction 5 mixture was acidified with glacial acetic acid to pH 4 and concentrated in vacuo. The residue was dissolved in a mixture of 200 ml ethyl acetate and 200 ml water. After additional washings
of the aqueous phase with ethyl acetate (3 x 50 ml) , the combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using ethyl acetate-hexane (1:4) as eluent to give the alcohol as a white solid (5.31 g) . MS: calc. monoisotopic mass: 351.2; found ESI+: 352.2
Synthesis of t-butyl- (3R, 4S) -4 - (N-benzyloxycarbonyl-N-methyl - amino) -3-methoxy-5-methyl-hexanoate
A solution of t-butyl- (3R, 4S) -4- (N-benzyloxycarbonylamino) -3- hydroxy- 5-methyl-hexanoate (3.027 g, 8.624 mmol) in tetrahydrofuran (40 ml) was added to a solution of LHMDS (24.0 mmol) in HMPA (4.5 ml, 25.7 mmol) and tetrahydrofuran (40 ml) at -78°C. After stirring for 20 min, methyltriflate (5.68 ml, 51.7 mmol) was added. After 1 h, the reaction was stopped by adding 70 ml aqueous 10% citric acid. The mixture was extracted with ethyl acetate (3x) . The combined organic extracts were washed with saturated aqueous NaHC03 (100 ml), dried over magnesium sulfate and concentrated in vacuo. The crude product was purified by column chromatography on silica gel, using ethyl acetate-hexane (1:4) as eluant to give the desired product as a colorless oil (2.456 g) . MS: calc. monoisotop. mass 379.24; found GAB-MS [M+H)+: 380
Synthesis of t-butyl- (3R, 4S) -4- (N-methylamino) -3-methoxy-5- methylhexanoate
To a solution of t-butyl- (3R,4S) -4- (N-benzyloxycarbonyl-N-methyl- amino) -3 -methoxy-5-methylhexanoate (3.855 g, 10.17 mmol) in
100 ml methanol was added 10% PD/C (0.541 g) and the mixture was hydrogenated until completion of the deprotection (tic control) . The catalyst was removed by filtration and the filtrate concentrated in vacuo. The resulting amorphous solid (2.49 g) can be crystallized from ether by adding a solution of HCl in dioxane. MS: calc. monoisotopic mass 245.2; found FAB-MS [M+H] + 246
Synthesis of t-butyl- (3R, 4S) -4- [N(benzyloxycarbonyl-L-valyl)N- methylamino] -3 -methoxy- 5 -me hylhexanoate
To a solution of Z-valine (3.672 g, 14.6 mmol) and pivaloyl chlorid (1.8 ml, 14.6 mmol) was added diisopropylethylamine (2.5 ml, 14.6 mmol) at -15°C. After 1 h, t-butyl- (3R, 4S) -4- (N- methylamino) -3 -methoxy-5-methylhexanoate (1.79 g, 7.3 mmol) and diisopropylethylamine (1.25 ml, 7.3 mmol) were added. The resulting mixture was stirred for 3 h at 0°C, 20 h at room temperature and then concentrated in vacuo. The residue was dissolved in
ethyl acetate (100 ml), washed with 10% aqueous citric acid (2 x 30 ml) and saturated aqueous NaHC03 (30 ml), dried over magnesium sulfate and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using ethyl acetate- hexane (1:4) as eluant to give the dipeptide as a colorless oil (1.804 g) . MS: calc. monoisotopic mass 478.3; found FAB-MS [M+H]+ 479
Synthesis of t-butyl - (3R, 4S) -4- [N(L-valyl) -N-methylamino) -3- methoxy- 5 -methylhexanoate
To a solution of t-butyl- (3R, S) -4- [N- (benzyloxycarbonyl-L- valyl) -N-methylamino) - -methoxy- 5 -methylhexanoate (1.804 g, 3.77 mmol) in methanol (60 ml) was added 10% Pd/C (0.26 g) and the mixture was hydrogenated until completion (tic control) . The catalyst was removed by filtration and the filtrate was concentrated in vacuo to give the deprotected dipeptide unit (1.27 g) . MS: calc. monoisotopic mass 344.27; found FAB-MS [M#H]+ 345
Synthesis of t-butyl- (3R, 4S) -4- [ (N,N-dimethyl-L-valyl-L-valyl) -N- methylamino) -3 -methoxy-5 -methyl -hexanoate
t-Butyl- (3R,4S) -4- [N- (L-valyl) -N-methylamino) -3 -methoxy- 5 -methylhexanoate (0.357 g, 1.038 mmol) and N,N-dimethylvaline (0.301 g, 2.076 mmol) were dissolved in 3 ml DMF and cooled to 0°C. DEPC (0.496 ml, 2.283 mmol) was added, followed by diisopropylethylamine (0.391 ml, 2.283 mmol). After stirring at 0°C for 3 h and at room temperature for 16 h, the reaction mixture was diluted with saturated aqueous NaHC03 (20 ml) was washed with toluene/ethyl - acetate 2:1 (3x) . The combined organic layers were extracted with 2 N NCI (3 x 10 ml). The aqueous phase was then neutralized with NaHC03 an extracted with toluene/ethyl acetate (2:1) (3 x 20 ml). The organic layer was dried over magnesium sulfate and concentrated in vacuo. The crude product was purified by crystalliza- tion from ether by adding a solution of HCl in dioxane to give the tripeptide as a white solid (0.433 g) . MS calc. monoisotopic mass 471.37; found FAB-MS [M+H] + 472
Synthesis of (3R, 4S) -4- [N- (N,N-dimethyl-L-valyl-L-valyl) -N- methylamino] -3-methoxy-5 -methylhexanoic acid
To a solution of t-butyl- (3R,4S) -4 - [N- (N,N-dimethyl-L-valyl-L- valyl) -N-methylamino] -3 -methoxy- 5 -methylhexanoate (0.22 g, 0,433 mmol) in dichloromethane (2 ml) was added trifluoroacetic acid (2 ml) . After stirring for 2 h, the reaction mixture was
concentrated in vacuo. Reevaporation of the residue with toluene (5 x 10 ml) gave the deprotected product (0.268 g) which was used for the next step without further purification. MS: calc. monoisotopic mass 415.3; found FAB-MS [M+H] + 416
Synthesis of N- (t-butyloxycarbonyl) -prolyl -prolyl -thiazolyl (2) - amide
Boc-prolyl-proline (1 g, 3.2 mmol) and 2-aminothiazol (0.357 g, 3.2 mmol) were dissolved in 25 ml DMF and cooled to 0°C. Triethyl- amine (1.1 ml, 7.6 mmol) was added, followed by DEPC (0.613 ml, 3.7 mmol) . After stirring at 0°C for 2 h and at room temperature for 24 h the reaction mixture was diluted with etyl acetate/ toluene (2:1), washed with 1 M aqueous potassium hydrogen sul- fate, water, saturated aqueous NaHC03 and aqueous NaCl , dried over sodium sulfate and concentrated in vacuo. The crude product was purified by crystallization from ethyl acetate (hexane to give a white solid (0.526 g) . Rf : 0.26 (ethyl acetate/hexane 1:1) ESI-MS: 395.2 [M+H] +, calc. C13H26 4θS = 394.3
Synthesis of prolyl -prolyl- thiazolyl (2) -amide
To a solution of N- (t-butoxycarbonyl) -propyl-propyl- thiazolyl (2) - amide in dichloromethane (2 ml) was added trifluoroacetic acid (2 ml) . After stirring for 2 h, the reaction mixture was concentrated in vacuo. Reevaporation of the residue with toluene (5 x 10 ml) gave the deprotected product which was used in the next step without further purification.
Synthesis of (3R, 4S) -4- [N- (N,N-dimethyl-L-valyl-L-valyl) -N- methylamino] -3 -methoxy-5-methyl -hexanoyl -prolyl-prolyl - thiazolyl (2) -amide (compound 1)
To a precooled solution of 0.1 g (3R,4S) -4- [N- [N,N-dimethyl-L- valyl-L-valyl) -N-methylamino] -3-methoxy-5-rnethylhexanoic acid (0.20 rnmol) and 0.08 g prolyl-prolyl- thiazolyl (2) -amide (0.20 mmol) in 1 ml DMF were added 0.037 ml DEPC (0.22 mmol) and 0.096 ml triethylamine (0.66 mmol). After stirring at 0°C for 2 h and at room temperature for 24 h, the reaction mixture was diluted with ethyl acetate/toluene (2:1), washed with 1 M aqueous potassium hydrogen sulfate, water, saturated aqueous NaHC03 and aqueous NaCl, dried over sodium sulfate and concentrated in vacuo. The crude product was purified via preparative HPLC to yield 112 mg of the desired product as a white solid. ESI -MS: 692 [M+H]+, calc. C34H57N706S = 691
Example 2 Synthesis Of (3R, 4S) -4 - [N- (N,N-dimethyl -L-valyl -L- valyl) -N-methylamino] -3 -methoxy-5 -methylhexanoyl- prolyl -prolyl -benzylamide (compound 2)
To a precooled solution of 0.15 g (3R, 4S) -4- [N- (N, N-dimethyl -L- valyl -L-valyl) -N-methylamino) -3-methoxy- 5-methyl -hexanoic acid (0.30 mmol) and 0.15 g prolyl-prolyl -benzylamide (0.30 mmol) in 2 ml DMF were added 0.2 ml DEPC (1.2 mmol) and 0.2 ml trietyl- amine (2.33 mmol). After stirring at 0°C for 2 h and at room temperature for 24 h, the reaction mixture was diluted with ethyl acetate/toluene (2:1), washed with 1 M aqueous potassium hydrogen sulfate, water, saturated aqueous NaHC03 and aqueous NaCl, dried over sodium sulfate and concentrated in vacuo. The crude product was purified via preparative HPLC to yield 161 mg of the desired product as a white solid.
ESI-MS: 699 [M+H] +, calc. C38H62N606 = 698
Example 3 Synthesis Of (3R, 4S, 5S) -4 - [N- (N,N-dimethyl-L-valyl-L- valyl) -N-methylamino] -3 -methoxy- 5 -methyl-heptanoyl- prolyl -prolyl -isopropylamide (compound 3)
Synthesis of N-Benzyloxycarbonyl -prolyl -isopropylamide
N-Benzyloxycarbonyl-proline (25 g, 0.1 mol) was dissolved in 450 ml dichloromethane. Then, 11.07 ml isopropylamine (0.13 mol) was added, followed by 7.66 g HOBt (0.05 mol), 24.92 g EDCI (0.13 mol) and 60.85 ml diisopropylethylamine (0.35 mol). The reaction mixture was stirred at room temperature overnight, further diluted with dichloromethane and washed with 5% citric acid (2x) , saturated sodium hydrogen carbonate (lx), and water (lx) . The organic layer was dried over sodium sulfate and concentrated in vacuo to yield 28.48 g of the product as an off-white solid. MS (EO) : M+ = 290.
Synthesis of Prolyl -isopropylamide
N-Benzyloxycarbonyl-prolyl-isopropylamide (28.48 g, 98.2 mmol) was dissolved in 75 ml methanol . This solution was added to a slurry of 3 spatulafuls of 10% Pd/C in 50 ml methanol under a stream of nitrogen. The reaction mixture was hydrogenated for 4 h, filtered over CELITE, washed with methanol, and the solvent evaporated under reduced pressure to give 16 g of a yellow crystalline solid. This material was dissolved in dichloromethane and washed with 5% citric acid (2x) . The combined acidic aqueous layers were brought to pH 12 with 1 N NaOH and solid NaOH, and
extracted with dichloromethane (2x) . The combined organic extracts were dried over sodium sulfate and concentrated in vacuo to yield 14.2 g of an off-white crystalline solid. MS (CI) : M+ = 157. 5
Synthesis of (3R,4S, 5S) -4- [N- (N,N-dimethyl-L-valyl-L-valyl) -N- methylamino) -3 -methoxy- 5 -methyl-heptanoyl -prolyl -prolyl -isopropylamide (compound 3)
10 (3R,4S,5S) -4- [N- {N,N-dimethyl-L-valyl -L-valyl) -N-methylamino) -3 -methoxy- 5 -methyl -heptanoyl -proline (0.45 g, 0.855 mmol) and 0.147 g prolyl- isopropylamide (0.940 mmol) were dissolved in 15 ml dichloromethane. After addition of 0.065 g HOBt (0.428 mmol), 0.18 g EDCI (0.94 mmol) and 0.44 g ml diiso-
15 propylethylamine. The reaction mixture was stirred at room temperature overnight. Additional reagents were added (0.073 g prolyl -isopropylamide, 0.032 g HOBt, 0.09 g ECI, 0.223 ml diisopropylethylamine) and stirring continued at room temperature overnight. The reaction mixture was concentrated in vacuo,
20 redissolved in ethyl acetate, and washed with saturated aqueous NaHC03, brine, and 5% citric acid. The acidic aqueous layers were brought to pH 10 with 1 N NaOH and extracted with ethyl acetate (2x) . The combined organic extracts were concentrated in vacuo. The residue was dissolved in 100 ml water and lyophilized to give
25 330 mg of the product as a fluffy white solid. ESI-MS: 665.5 [M+H]+
The following compounds can be prepared according to the methods described in Example 1, 2, and 3:
30
COMPOUND COMPOUND SEQUENCE
66 Xaa Val Xad Pro Xaw
67 Xaa Val Xad Pro Xax
68 Xaa Val Xad Pro Xay
69 Xaa Val Xad Pro Xba 5 70 Xaa Val Xad Pro Xbb
71 Xaa Val Xad Pro Xbc
72 Xaa Val Xad Pro Xbd
73 Xaa Val Xad Pro Xbe
74 Xaa Val Xad Pro Xbf 10 75 Xaa Val Xad Pro Xbg
76 Xaa Val Xad Pro Xbh
77 Xaa Val Xad Pro Xbi
78 Xaa Val Xad Pro Xbk
79 Xaa Val Xad Pro Xbl 15 80 Xaa Val Xad Pro Xbm
81 Xaa Val Xad Pro Xbn
82 Xaa Val Xad Pro Xbo
83 Xaa Val Xad Pro Xbp
84 Xaa Val Xad Pro Xbq 20 85 Xaa Val Xad Pro Xbr
86 Xaa Val Xad Pro Xbs
87 Xaa Val Xad Pro Xbt
88 Xaa Val Xad Pro Xbv
89 Xaa Val Xad Pro Xbw 25 90 Xaa Val Xad Pro Xbx
91 Xab Val Xae Pro Xbo
92 Xae Val Xae Pro Xbo
93 Xaa He Xae Pro Xbo
94 Xaa Xbu Xae Pro Xbo 30 95 Xab Val Xad Pro Xbo
96 Xae Val Xad Pro Xbo
97 Xaa He Xad Pro Xbo
98 Xaa Xbu Xad Pro Xbo
99 Xaa Val Xae Pro Xbz 35
The symbols X in the summary have the following meanings:
Xaa: N,N-Dimethyl-valine 0
Xab: N,N-Dimethylisoleucine
c Xae: N,N-Dimethyl-tert-leucine
H3C
CH3
H3C
0 H3C
H3C
CHs
Xbu: 2-tert,butylglycine
Xby: prolyl tert .butylester
Xbz: prolyl benzylamide
Example 4 Determination of In Vitro Cytotoxicity
Cytotoxicity was measured using the microculture tetrazolium assay (MTT) , a standard methodology for adherent cell lines. Details of this assay have been published (Alley, et al.. Cancer Research 48: 589-601 (1988)). Exponentially growing cultures of HT-29 colon carcinoma cells were used to make microtiter plate cultures. Cells were seeded at 5000-20,000 cells per well in 96-well plates (in 150 μl of media), and grown overnight at 37°C. Test compounds were added in 10-fold dilutions varying from 10 " to 10-ι° M. Cells were then incubated for 48 hours. To determine the number of viable cells in each well, the MTT dye was added
(50 μl of 3 mg/ml solution of 3- (4, 5-dimethylthiazol -2 -yl) -2, 5-di- phenyltetrazolium bromide in saline) . This mixture was incubated
at 37°C for 5 hours, and then 50 μl of 25% SDS, pH 2, was added to each well. After an overnight incubation, the absorbance of each well at 550 nm was read using an ELISA reader. The values for the mean +/ - SD of data from replicated wells were calcu- lated, using the formula % T/C (% viable cells treated/control) .
OD of treated cells x 100 = % T/C
OD of control cells
The concentration of test compound which gives a T/C of 50% growth inhibition was designated as the IC50 value.
Results
The results of the in vitro evaluation are shown in the table below. The IC50 values shown are in the nanomolar range, indicating that these compounds possess significant activity in the HT-29 system.
Table
COMPOUND IC50 (M)
1 2 x IO-9 2 4 x lO"9
3 6 x lO"9
Example 5 Determination of In Vivo Activity
Compounds of this invention can be tested in a pre- clinical assay for in vivo activity which is indicative of clinical utility. Such assays are typically conducted with nude mice into which tumor tissue, preferably of human origin, had been transplanted (xenografted) , as is well known in this field. Test compounds are evaluated for anti -tumor efficacy following administration to the xenograft-bearing mice.
For example, human breast tumors (MX-1) which have been growth in athymic nude mice are transplanted into new recipient mice, using tumor fragments of about 50 mg in size. The day of transplantation is designated as day 0. The mice are divided into four groups of 5-10 mice each. An untreated group serves as the control. Doses can be, for example, administered on a schedule such as on days 5, 7, 9, 12, 14, 16, 19, 21 and 23 post -implantation.
Tumor diameters and body weights are measured twice weekly. Tumor volumes are calculated using the diameters measured with Vernier calipers, and the formula
(length x width2) /2 = mm3 of tumor volume
Mean tumor volumes are calculated for each treatment group, and T/C values determines for each group relative to the untreated control tumors.
In another in vivo assay, the P388 murine lymphocytic leukemia is harvested from donor mice by peritoneal lavage at day 7 post- transplant and administered to test mice. Treatment of test mice begins 3 days post-transplant and the drugs are administered for five consecutive days. The survival time for untreated mice is typically between 11 and 13 days. The data are expressed as mean survival time (MST) and tretade/control %. According to National Cancer Institute guidelines, a T/C in the range of 128-190 % indicates a drug with moderate to good activity.
Equivalents
Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments are intended to be encompassed in the scope of the following claims.
Claims
1 . A compound of Formula I ,
A-B-N(CH3) -CHD-CH(OCH3) -CH2CO-Pro- Pro- K (I) ,
wherein A is an amino acid residue of the formula
(CH3)2N-CHX-CO, wherein X is a normal or branched C3-C -alkyl group; B is an amino acid residue selected from the group consisting of valyl, isoleucyl, leucyl, and 2 -t-butylglycyl; D is a normal or branched C2-Cs-alkyl group; and K is a t-butoxy group or a substituted amino group; or a salt thereof with a pharmaceutically acceptable acid.
2. The compound of Claim 1 wherein K is a substituted amino group selected from the group consisting of -N(Cι-3-alkyl)Cι-3-alkyl, normal or branched -NH-C1-8-alkyl, -NH-C(CH3)2CN, -NH-C(CH3)2CCH, -NH-C (CH3 ) 2CH2CH2OH, -NH-C(CH3)2CH2OH, -NH-C3-8.cycloalkyl , -NH- [3, 3, 0] -bicyclooctyl, -NHCH(CH3)CH(OH)C6H5, -NH-quinolyl, -NH-pyrazyl, -NH-CH2-benzimidazolyl, -NH-adamantyl, -NH-CH -adamantyl, -NH-CH(CH3) -phenyl, -NH-C (CH3) 2-phenyl, -NfC^-alkoxy) -Cι-4- alkyl, N (Cι- -alkoxy) -CH2-phenyl, -N(C_.-4-alkoxy) phenyl, -N(CH3)0-phenyl, -NH- (CH2)v-phenyl (v = 0, 1, 2, or 3), -NH- (CH2)m-naphthyl (m = 0 or 1) , -NH- (CH2)w-benzhydryl (w = 0, 1, or 2), -NH-biphenyl, -NH-pyridyl, -NH-CH2-pyridyl, -NH-CH -CH2-pyridyl, -NH-benzothiazolyl, -NH-benzoisothia- zolyl, -NH-benzopyrazolyl, -NH-benzoxazolyl, -NH- (CH2)m-fluor- enyl (m = 0 or 1) , -NH-pyrimidyl, -NH- <CH2)m-in.danyl (m = 0 or 1), -NH- (CH2CH20)v-CHCH3 (y = 0, 1, 2, 3, 4, or 5), -NH- (CH2CH20)y-CH3 (y = 0, 1, 2, 3, 4, or 5), -NH-NH-C6H5, -NH-N(CH3)C6H5, -NH-NH-CH2-C6H5, -NH-N (CH3) CH2-C6H5,
H3C
3. The compound of Claim 1 wherein X is an isopropyl, t-butyl or sec -butyl group; B is valyl, isoleucyl, or 2- t-butylglycyl; D is an isopropyl, t-butyl or sec -butyl group; and K is a t-butoxy group or a substituted amino group.
4. The compound of Claim 3 wherein K is a substituted amino group selected from the group consisting of -NHCH3, -NHCHCH3, -NH(CH2)2CH3, -NH(CH2)3CH3, -NH (CH2) CH3 , -NH (CH2) 5CH3 , -NHCH(CH3)2, -NHCH(CH2CH3)2, -NHCH (CH2CH2CH3) 2, -NHC(CH3)3, -NHCH[CH(CH3)2-]2, -NHCH (CH2CH3) CH2CH2CH3, -NHCH (CH3)CH2CH3, -NHCH2CH2F, -NHC(CH3)2CH2CH3, -NHCH (CH3)CH (CH3) 2, -NHCH(CH3)C(CH3)3, -NHCH(CH3) CH2CH2CH3, -NHCH2CH (CH2) 2, -NHCH2C(CH3)3, -NH-cyclopropyl, -NH-cyclobutyl, -NH-cyclo- pentyl, -NH-cyclohexyl, -NH-cycloheptyl, -N(CH3)0CH3, -N(CH3)2, -N(CH3)OCH2CH3, -N (CH3)OCH2CHCH3, -N(CH3) OCH (CH3) 2, -N(CH2CH3)OCH3, -N(CH2CH3)OCH2CH3, -N(CH3) OCH2C6H5, -N(OCH3)CH2-C6H5, N(CH3)OC6H5, -NH-CH2-C€H5, -NH (CH2) 2C6H5, -NH{CH2)3C6H5, -NHCH(CH3)CH(OH)C6H5, -NH-CH2-cyclohexyl , -NH-indanyl- (1) , -NH-CH2CF3, -NHCH(CH2F)2, -NHC (CH3) 2CH2OH, -NH(CH2CH20)2CH2CH3, -NHC (CH3) CN, -NH-quinolyl, -Nh-pyrazyl, -NH-adamantyl (2) , -NH-adamantyl (1) , -NH-CH -naphthyl, -NH-benzhydryl, -NH-biphenyl, -NH-pyridyl, -NH-CH2pyridyl, -NH-CH2-CH2-pyridyl, -NH-benzothiazolyl, -NH-benzoisothia- zolyl, -NH-benzopyrazolyl, -NH-benzoxazolyl, -NH-fluorenyl, -NH-pyrimidyl, -NH-CH2- (4 -methyl) thiazolyl (2) ,
-NH-CH2-furanyl(2) , -NH-CH2- thienyl (2) , -NH-CH2- (5-methyl) - thienyl(2), -NH- thiazolyl (2) , -NH-isoxazolyl (3) , -NH- (3 -methyl) isoxazolyl (5) , -NH- (3 -methyl) isothiazolyl (5) , -NH- (5 -trifluoromethyl) thiadiazolyl (2) , -NH- (5-cyclo- propyl) thiadiazolyl (2) , -NH- (4, 5 -dimethyl) thiazolyl (2) , -NH- (5-methyl) thiadiazolyl (2) ,
..j /—\
—N -N S=0 -N v_7
■N . —N 0 and —N s vJ _7 \_7 ^0
5. The compound of Claim 3 wherein K is a substituted amine selected from the group consisting of -NHCH3, -N(CH3)2, -NH(CH2)2CH3, -NHCH(CH3)2, -NHCH [CH(CH3) 2] 2, *NHCH(CH3) CH2CH3, -NH(CH2)5CH3, -NHCH(CH2CH3)2, -NHCH (CH3) 2CH2CH3, -NHCH(CH2CH2CH2)2, -NHC(CH3)3, -NHCH(CH2CH3)CH2CH2CH2, -NHCH(CH3)C(CH3)3, -NHCH2CH(CH3) 2, -NHCH2C (CH3) 3 , -NH-cyclO- propyl, -NH-eye1opentyl, -NH-cyclohexyl, -NH-cycloheptyl, -N (CH3 ) OCH3 , -N (CH3) (0CH2CH3) , -N (CH3) OCH2C6H5, -NH-CH(CH3) -C6H6, -NHCH2CH2C6H5, - NHCH (CH3 ) CH (OH)C6H5, -NH-CH2-cyclohexyl, -NH- (CH2CH20) 2CH2CH3, -NH- indanyl- (1) , -NHCH(CH2F)2, -NHC(CH3)2CN, -NH-CH2 -CH2 -pyridyl , 4 -morpholinyl, 2 - thiazolidinyl, and -NH- (5-methyl) thia- diazolyl(2).
6. The compound of Claim 5 wherein X is isopropyl, B is valyl, and D is isopropyl or sec-butyl.
7. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of Claim 1.
8. A method for treating a tumor in a mammal, comprising administering to the mammal a tumor- inhibiting amount of a compound of Claim 1.
9. The method of Claim 10 wherein the tumor is a colon tumor, a breast tumor or a lung tumor.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US688334 | 1996-07-30 | ||
US08/688,334 US5741892A (en) | 1996-07-30 | 1996-07-30 | Pentapeptides as antitumor agents |
PCT/EP1997/003902 WO1998004581A1 (en) | 1996-07-30 | 1997-07-21 | Dolastatin pentapeptide derivatives and their use as antitumor agents |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0915908A1 true EP0915908A1 (en) | 1999-05-19 |
Family
ID=24764016
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP97934502A Withdrawn EP0915908A1 (en) | 1996-07-30 | 1997-07-21 | Dolastatin pentapeptide derivatives and their use as antitumor agents |
Country Status (7)
Country | Link |
---|---|
US (1) | US5741892A (en) |
EP (1) | EP0915908A1 (en) |
JP (1) | JP2000516923A (en) |
AU (1) | AU3769297A (en) |
TW (1) | TW472063B (en) |
WO (1) | WO1998004581A1 (en) |
ZA (1) | ZA976725B (en) |
Families Citing this family (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DK0831100T3 (en) * | 1995-04-21 | 2001-01-02 | Teikoku Hormone Mfg Co Ltd | New peptide derivatives |
US6323315B1 (en) | 1999-09-10 | 2001-11-27 | Basf Aktiengesellschaft | Dolastatin peptides |
CA2385528C (en) | 1999-10-01 | 2013-12-10 | Immunogen, Inc. | Compositions and methods for treating cancer using immunoconjugates and chemotherapeutic agents |
US6884869B2 (en) * | 2001-04-30 | 2005-04-26 | Seattle Genetics, Inc. | Pentapeptide compounds and uses related thereto |
US7256257B2 (en) | 2001-04-30 | 2007-08-14 | Seattle Genetics, Inc. | Pentapeptide compounds and uses related thereto |
EP1545613B9 (en) | 2002-07-31 | 2012-01-25 | Seattle Genetics, Inc. | Auristatin conjugates and their use for treating cancer, an autoimmune disease or an infectious disease |
KR101520209B1 (en) | 2003-11-06 | 2015-05-13 | 시애틀 지네틱스, 인크. | Monomethylvaline compounds capable of conjugation to ligands |
US7853332B2 (en) * | 2005-04-29 | 2010-12-14 | Medtronic, Inc. | Lead electrode for use in an MRI-safe implantable medical device |
WO2006122408A1 (en) | 2005-05-18 | 2006-11-23 | Aegera Therapeutics Inc. | Bir domain binding compounds |
US8163792B2 (en) | 2006-05-16 | 2012-04-24 | Pharmascience Inc. | IAP BIR domain binding compounds |
TW200918089A (en) | 2007-07-16 | 2009-05-01 | Genentech Inc | Humanized anti-CD79b antibodies and immunoconjugates and methods of use |
JP5469600B2 (en) | 2007-07-16 | 2014-04-16 | ジェネンテック, インコーポレイテッド | Anti-CD79b antibody and immunoconjugate and method of use thereof |
SI2657253T1 (en) | 2008-01-31 | 2017-10-30 | Genentech, Inc. | Anti-CD79b antibodies and immunoconjugates and methods of use |
ES2625637T3 (en) | 2010-02-12 | 2017-07-20 | Pharmascience Inc. | BIR IAP domain binding compounds |
US10131682B2 (en) | 2012-11-24 | 2018-11-20 | Hangzhou Dac Biotech Co., Ltd. | Hydrophilic linkers and their uses for conjugation of drugs to a cell binding molecules |
US10464955B2 (en) | 2014-02-28 | 2019-11-05 | Hangzhou Dac Biotech Co., Ltd. | Charged linkers and their uses for conjugation |
PL3262071T3 (en) | 2014-09-23 | 2020-08-10 | F. Hoffmann-La Roche Ag | Method of using anti-cd79b immunoconjugates |
CN113350518A (en) | 2015-07-12 | 2021-09-07 | 杭州多禧生物科技有限公司 | Conjugated bridge linkers to cell binding molecules |
US9839687B2 (en) | 2015-07-15 | 2017-12-12 | Suzhou M-Conj Biotech Co., Ltd. | Acetylenedicarboxyl linkers and their uses in specific conjugation of a cell-binding molecule |
NZ752394A (en) | 2016-11-14 | 2021-07-30 | Hangzhou Dac Biotech Co Ltd | Conjugation linkers, cell binding molecule-drug conjugates containing the likers, methods of making and uses such conjugates with the linkers |
CA3230737A1 (en) | 2021-09-03 | 2023-03-09 | Toray Industries, Inc. | Pharmaceutical composition for cancer treatment and/or prevention |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4816444A (en) * | 1987-07-10 | 1989-03-28 | Arizona Board Of Regents, Arizona State University | Cell growth inhibitory substance |
US4879278A (en) * | 1989-05-16 | 1989-11-07 | Arizona Board Of Regents | Isolation and structural elucidation of the cytostatic linear depsipeptide dolastatin 15 |
DE69230824T2 (en) * | 1991-08-09 | 2000-07-27 | Teikoku Hormone Mfg Co Ltd | NEW TETRAPEPTIDE DERIVATIVES |
CA2136339C (en) * | 1992-05-20 | 2008-02-05 | Andreas Haupt | Derivatives of dolastatin |
NZ258882A (en) * | 1992-12-16 | 1997-06-24 | Basf Ag | Antineoplastic peptides and compositions thereof |
US5504191A (en) * | 1994-08-01 | 1996-04-02 | Arizona Board Of Regents Acting On Behalf Of Arizona State University | Human cancer inhibitory pentapeptide methyl esters |
US5530097A (en) * | 1994-08-01 | 1996-06-25 | Arizona Board Of Regents Acting On Behalf Of Arizona State University | Human cancer inhibitory peptide amides |
US5663149A (en) * | 1994-12-13 | 1997-09-02 | Arizona Board Of Regents Acting On Behalf Of Arizona State University | Human cancer inhibitory pentapeptide heterocyclic and halophenyl amides |
-
1996
- 1996-07-30 US US08/688,334 patent/US5741892A/en not_active Expired - Fee Related
-
1997
- 1997-07-21 EP EP97934502A patent/EP0915908A1/en not_active Withdrawn
- 1997-07-21 AU AU37692/97A patent/AU3769297A/en not_active Abandoned
- 1997-07-21 WO PCT/EP1997/003902 patent/WO1998004581A1/en not_active Application Discontinuation
- 1997-07-21 JP JP10508458A patent/JP2000516923A/en active Pending
- 1997-07-29 ZA ZA976725A patent/ZA976725B/en unknown
- 1997-07-30 TW TW086110885A patent/TW472063B/en not_active IP Right Cessation
Non-Patent Citations (1)
Title |
---|
See references of WO9804581A1 * |
Also Published As
Publication number | Publication date |
---|---|
ZA976725B (en) | 1999-02-12 |
AU3769297A (en) | 1998-02-20 |
JP2000516923A (en) | 2000-12-19 |
US5741892A (en) | 1998-04-21 |
WO1998004581A1 (en) | 1998-02-05 |
TW472063B (en) | 2002-01-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US5741892A (en) | Pentapeptides as antitumor agents | |
EP0831100B1 (en) | Novel peptide derivatives | |
EP0871656B1 (en) | Dolastatin derivatives, their preparation and use | |
US6323315B1 (en) | Dolastatin peptides | |
US8163698B2 (en) | Dolastatin 15 derivatives | |
EP0674652B1 (en) | Dolastatin analog | |
WO1998040400A1 (en) | Dolastatin 15 derivatives with carbonyl and heterocyclic functionalities at the c-terminus | |
RU2116312C1 (en) | Peptide derivatives or their salts, pharmaceutical composition | |
US5939527A (en) | Tetrapeptides as antitumor agents | |
EP0859786A1 (en) | Peptide derivatives of dolastatin 15 and their use | |
EP1593686B1 (en) | Antineoplastic peptides | |
AU775090B2 (en) | Antineoplastic peptides | |
SK282467B6 (en) | Peptides, pharmaceutical preparation containing them and their use |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 19981217 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): CH DE FR GB IT LI NL |
|
17Q | First examination report despatched |
Effective date: 20000816 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20020903 |